Characterization of a comprehensive two-dimensional anion exchange-perfusive reversed phase liquid chromatography system for improved separations of peptides

Improved two-dimensional separations of nanoliter volumes of peptides are presented. The two-dimensional anion exchange-reversed phase chromatography system described here is the result of several modifications of a two-dimensional system described previously. The primary change is the implementation of a perfusive stationary phase in the second separation dimension. The improved system has a peak capacity of 2028 as compared to 1050 for the previously described system. The limit of detection is 0.78 attomoles. The higher resolving power and retention time reproducibility of the modified system are demonstrated with two-dimensional chromatograms of the peptides obtained from a tryptic digest of porcine thyroglobulin. © 2000 John Wiley & Sons, Inc. J Micro Sep 12: 371–377, 2000